Tlhahlobo e fapaneng ea li-nodule tsa pulmonary tse khetholloang ke computed tomography (CT) e ntse e le phephetso ts'ebetsong ea bongaka.Mona, re tšoaea metabolome ea lefats'e ea lisampole tsa serum tse 480, ho kenyelletsa le taolo e phetseng hantle, maqhutsu a mats'oafo, le sethala sa I lung adenocarcinoma.Adenocarcinomas e bonts'a lintlha tse ikhethang tsa metabolomic, athe li-nodule tse nang le bokooa le batho ba phetseng hantle ba na le ho tšoana ho hoholo ho profil ea metabolomic.Sehlopheng sa ho sibolla (n = 306), sete sa metabolite ea 27 se ile sa khetholloa ho khetholla pakeng tsa li-nodule tse kotsi le tse kotsi.AUC ea mohlala oa khethollo ho netefatso ea ka hare (n = 104) le lihlopha tsa ka ntle (n = 111) e ne e le 0.915 le 0.945, ka ho latellana.Tlhahlobo ea tsela e senotse ho eketseha ha metabolites ea glycolytic e amanang le ho fokotseha ha tryptophan serum ea matšoafo adenocarcinoma ha e bapisoa le maqhutsu a nang le bokooa le taolo e phetseng hantle, mme a fana ka tlhahiso ea hore ho nka tryptophan ho khothalletsa glycolysis liseleng tsa mofetše oa matšoafo.Boithuto ba rona bo totobatsa boleng ba li-biomarker tsa serum metabolite ho lekola kotsi ea li-nodule tsa pulmonary tse fumanoeng ke CT.
Ho hlahlojoa esale pele ho bohlokoa ho ntlafatsa maemo a bophelo ho bakuli ba mofetše.Liphello tse tsoang ho US National Lung Cancer Screening Trial (NLST) le Thuto ea NELSON ea Europe li bontšitse hore ho hlahloba ka tekanyo e tlaase ea computed tomography (LDCT) ho ka fokotsa haholo lefu la kankere ea matšoafo lihlopheng tse kotsing e kholo1,2,3.Ho tloha ha ts'ebeliso e atileng ea LDCT bakeng sa tlhahlobo ea mofets'e oa matšoafo, ts'ebetso ea liphuputso tsa radiographic tsa liketsahalo tsa asymptomatic pulmonary nodules li tsoetse pele ho eketseha 4.Li-nodule tsa matšoafo li hlalosoa e le li-opacities tse tsepamisitsoeng ho fihlela ho 3 cm ka bophara ba 5.Re tobane le mathata a ho lekola monyetla oa lefu le ho sebetsana le palo e kholo ea maqhutsu a matšoafo a fumanoeng ka tšohanyetso ho LDCT.Meeli ea CT e ka lebisa litlhahlobong tsa ho latela khafetsa le liphetho tse fosahetseng, tse lebisang ho keneng ho sa hlokahaleng le kalafong e feteletseng6.Ka hona, ho na le tlhokahalo ea ho hlahisa li-biomarker tse ka tšeptjoang le tse molemo ho khetholla kankere ea matšoafo ka nepo methating ea pele le ho khetholla boholo ba li-nodule tse nang le bokooa ha ho fumanoa qalong 7.
Tlhahlobo e felletseng ea limolek'hule tsa mali (serum, plasma, peripheral blood mononuclear cell), ho kenyelletsa le genomics, proteomics kapa DNA methylation8,9,10, e lebisitse thahasellong e ntseng e hola ea ho sibolloa ha li-biomarker tsa tlhahlobo ea mofetše oa matšoafo.Ho sa le joalo, mekhoa ea metabolomics e lekanya lihlahisoa tsa lisele tsa cellular tse susumetsoang ke liketso tsa endogenous le exogenous 'me ka hona li sebelisoa ho bolela esale pele ho qala ha lefu le phello.Liquid chromatography-tandem mass spectrometry (LC-MS) ke mokhoa o sebelisoang haholo bakeng sa lithuto tsa metabolomics ka lebaka la kutloisiso ea eona e phahameng le mefuta e mengata e matla, e ka koahelang metabolites e nang le mefuta e fapaneng ea physicochemical properties11,12,13.Le hoja tlhahlobo ea lefats'e ea metabolite ea plasma / serum e 'nile ea sebelisoa ho khetholla li-biomarker tse amanang le ho hlahlojoa ha kankere ea matšoafo14,15,16,17 le katleho ea phekolo, lihlopha tse 18 tsa serum metabolite ho khetholla pakeng tsa li-nodule tse kotsi le tse kotsi tsa matšoafo li ntse li tsoela pele ho ithutoa haholo.- lipatlisiso tse kholo.
Adenocarcinoma le squamous cell carcinoma ke mefuta e 'meli ea mantlha ea mofets'e o seng o monyane oa mofetše oa matšoafo oa sele (NSCLC).Liteko tse fapaneng tsa tlhahlobo ea CT li bonts'a hore adenocarcinoma ke mofuta o atileng haholo oa mofets'e oa matšoafo1,19,20,21.Thutong ena, re sebelisitse ultra-performance liquid chromatography-high-resolution mass spectrometry (UPLC-HRMS) ho etsa tlhahlobo ea metabolomics ka kakaretso ea lisampole tsa serum tse 695, ho kenyeletsoa li-control tse phetseng hantle, li-nodule tsa benign pulmonary, le CT-tse fumanoeng ≤3 cm.Ho hlahlojoa ha Stage I lung adenocarcinoma.Re khethile sehlopha sa serum metabolites se khethollang adenocarcinoma ea matšoafo ho tloha maqhubung a bobebe le taolo e phetseng hantle.Tlhahlobo ea ntlafatso ea tsela e senotse hore tryptophan e sa tloaelehang le metabolism ea glucose ke liphetoho tse tloaelehileng ho lung adenocarcinoma ha li bapisoa le maqhutsu a nang le bokooa le taolo e phetseng hantle.Qetellong, re thehile le ho netefatsa sehlopha sa metabolic sa serum se nang le maemo a holimo le kutlo ho khetholla lipakeng tsa maqhutsu a mabe le a bobebe a fumanoeng ke LDCT, a ka thusang tlhahlobong ea pele ea phapang le tlhahlobo ea kotsi.
Phuputsong ea morao-rao, lisampole tsa serum tse tsamaellanang le thobalano le lilemo li ile tsa bokelloa ka morao ho tloha ho li-control tse 174 tse phetseng hantle, bakuli ba 292 ba nang le maqhutsu a mats'oafo, le bakuli ba 229 ba nang le sethala sa I lung adenocarcinoma.Litšobotsi tsa palo ea batho ba lithuto tse 695 li bontšoa ho Lethathamo la 1 la Tlatsetso.
Joalokaha ho bontšitsoe Figure 1a, kakaretso ea 480 disampole serum, ho akarelletsa le 174 ho laola bophelo bo botle (HC), 170 benign nodules (BN), le 136 sethaleng I lung adenocarcinoma (LA) disampole, li ile tsa bokelloa ka Sun Yat-sen University Cancer Center.Discovery cohort bakeng sa profiling e sa rereloang ea metabolomic e sebelisang ultra-performance liquid chromatography-high-resolution mass spectrometry (UPLC-HRMS).Joalokaha ho bontšitsoe ho Setšoantšo sa Tlatsetso ea 1, ho ile ha khethoa metabolite e fapaneng pakeng tsa LA le HC, LA le BN ho theha mohlala oa lihlopha le ho hlahloba ho feta tlhahlobo ea litsela tse fapaneng.Mehlala ea 104 e bokelletsoeng ke Setsi sa Kankere sa Univesithi ea Sun Yat-sen le lisampole tse 111 tse bokelletsoeng ke lipetlele tse ling tse peli li ile tsa netefatsoa ka hare le kantle, ka ho latellana.
palo ea batho ba ithutoang sehlopheng sa ho sibolla se entseng tlhahlobo ea serum metabolomics ea lefats'e ho sebelisa ultra-performance liquid chromatography-high-resolution mass spectrometry (UPLC-HRMS).b Tlhahlobo e sa lekanyetsoang ea lisekoere tse nyane (PLS-DA) ea kakaretso ea metabolome ea 480 ea lisampole tsa serum ho tsoa sehlopheng sa boithuto, ho kenyeletsoa le taolo e phetseng hantle (HC, n = 174), maqhutsu a nang le bokooa (BN, n = 170), le sethala sa I lung adenocarcinoma. (Los Angeles, n = 136).+ ESI, mokhoa o motle oa ionization oa electrospray, -ESI, mokhoa o fosahetseng oa ionization oa electrospray.c–e Metabolites e nang le bongata bo fapaneng haholo lihlopheng tse peli tse fanoeng (teko ea maemo a saenneng ea Wilcoxon ea mehatla e 'meli, sekhahla sa ho sibolloa ha leshano se fetotsoe boleng ba p, FDR <0.05) e bonts'itsoe ka bofubelu (phetoho ea mena> 1.2) le boputsoa (phetoho <0.83) .) e bonts'itsoeng setšoantšong sa seretse se chesang.f 'Mapa oa mocheso oa sehlopha sa Hierarchical o bonts'ang phapang e kholo palong ea metabolite e hlalositsoeng lipakeng tsa LA le BN.Lintlha tsa mohloli li fanoa ka mokhoa oa lifaele tsa data tsa mohloli.
Kakaretso ea serum metabolome ea 174 HC, 170 BN le 136 LA sehlopheng sa ho sibolla e ile ea hlahlojoa ho sebelisoa tlhahlobo ea UPLC-HRMS.Re qala ka ho bonts'a hore lisampole tsa taolo ea boleng (QC) li kopane ka thata bohareng ba mohlala oa principal component analysis (PCA) o sa laoleheng, o tiisang botsitso ba mosebetsi oa hona joale oa thuto (Supplementary Figure 2).
Joalo ka ha ho bonts'itsoe tlhahlobong e sa lekanyetsoang ea lisekoere-khethollo (PLS-DA) setšoantšong sa 1 b, re fumane hore ho na le phapang e hlakileng lipakeng tsa LA le BN, LA le HC ka mekhoa e metle (+ESI) le e mpe (−ESI) ea ionization ea electrospray. .ka thoko.Leha ho le joalo, ha ho phapang e kholo e fumanoeng pakeng tsa BN le HC maemong a +ESI le -ESI.
Re fumane likarolo tse 382 tse fapaneng pakeng tsa LA le HC, likarolo tse 231 tse fapaneng pakeng tsa LA le BN, le likarolo tse 95 tse fapaneng pakeng tsa BN le HC (teko ea boemo bo saenneng ea Wilcoxon, FDR <0.05 le liphetoho tse ngata> 1.2 kapa <0.83) (Figure .1c-e) )..Litlhōrō li ile tsa hlakisoa ka ho eketsehileng (Supplementary Data 3) khahlanong le database (mzCloud/HMDB/Chemspider library) ka boleng ba m/z, nako ea ho boloka le ho batla ho arohana ha bongata (lintlha tse hlalositsoeng karolong ea Mekhoa) 22.Qetellong, 33 le 38 e hlalosang metabolites e nang le phapang e khōlō ka bongata e ile ea khetholloa bakeng sa LA khahlanong le BN (Setšoantšo sa 1f le Supplementary Table 2) le LA khahlanong le HC (Supplementary Figure 3 le Supplementary Table 2), ka ho latellana.Ka lehlakoreng le leng, ke li-metabolite tsa 3 feela tse nang le phapang e kholo ea bongata tse ileng tsa khetholloa ho BN le HC (Supplementary Table 2), e lumellanang le ho kopana pakeng tsa BN le HC ho PLS-DA.Li-metabolite tsena tse fapaneng li koahela mefuta e mengata ea lik'hemik'hale tsa likokoana-hloko (Supplementary Figure 4).Ha li kopane, liphetho tsena li bonts'a liphetoho tse kholo ho serum metabolome e bonts'ang phetoho e mpe ea mofets'e oa pele oa matšoafo ha o bapisoa le maqhutsu a mats'oafo kapa batho ba phetseng hantle.Ho sa le joalo, ho tšoana ha serum metabolome ea BN le HC ho fana ka maikutlo a hore li-nodule tsa benign pulmonary li ka arolelana litšobotsi tse ngata tsa likokoana-hloko le batho ba phelang hantle.Ka lebaka la hore liphetoho tsa liphatsa tsa lefutso tsa epidermal growth factor receptor (EGFR) li tloaelehile ho lung adenocarcinoma subtype 23, re ile ra batla ho fumana hore na liphetoho tsa mokhanni li ama serum metabolome.Ka mor'a moo re ile ra hlahloba boemo ba kakaretso ba metabolomic ea linyeoe tsa 72 tse nang le boemo ba EGFR sehlopheng sa lung adenocarcinoma.Hoa thahasellisa hore re fumane litlaleho tse bapisoang pakeng tsa bakuli ba EGFR ba feto-fetohang (n = 41) le EGFR bakuli ba mofuta o hlaha (n = 31) tlhahlobo ea PCA (Supplementary Figure 5a).Leha ho le joalo, re ile ra khetholla metabolites ea 7 eo bongata ba bona bo ileng ba fetoha haholo ho bakuli ba nang le phetoho ea EGFR ha ba bapisoa le bakuli ba nang le mofuta oa hlaha oa EGFR (teko ea t, p <0.05 le phetoho ea fold> 1.2 kapa <0.83) (Supplementary Figure 5b).Bongata ba metabolite tsena (5 ho tse 7) ke li-acylcarnitine, tse bapalang karolo ea bohlokoa litseleng tsa oxidation ea mafura a acid.
Joalokaha ho bontšitsoe setšoantšong sa 2 a, li-biomarker bakeng sa lihlopha tsa li-nodule li ile tsa fumanoa ho sebelisoa basebetsi ba fokolang ka ho feletseng le khetho e thehiloeng ho li-metabolite tse fapaneng tsa 33 tse khetholloang ho LA (n = 136) le BN (n = 170).Motsoako o motle ka ho fetisisa oa mefuta-futa (LASSO) - binary logistic regression model.Ho ile ha sebelisoa ho netefatsoa ha sefapano ka makhetlo a leshome ho leka ho tšepahala ha mohlala.Khetho e feto-fetohang le ts'ebetso ea paramethara e lokisoa ke kotlo ea ho eketsa monyetla ka paramente λ24.Tlhahlobo ea lefats'e ea metabolomics e ile ea tsoela pele ho etsoa ka boithaopo ho netefatso ea ka hare (n = 104) le lihlopha tsa ho netefatsa ka ntle (n = 111) ho hlahloba ts'ebetso ea lihlopha tsa mohlala oa khethollo.Ka lebaka leo, li-metabolite tsa 27 ha li sibolloa li ile tsa khetholloa e le mohlala o motle ka ho fetisisa oa khethollo o nang le moelelo o moholo ka ho fetisisa oa AUC (setšoantšo sa 2b), eo 9 e neng e e-na le ts'ebetso e eketsehileng le 18 e fokotsehile ho LA ha e bapisoa le BN (setšoantšo sa 2c).
Phallo ea mosebetsi bakeng sa ho aha sehlopha sa li-nodule tsa pulmonary, ho kenyelletsa le ho khetha karolo e ntle ka ho fetisisa ea metabolites ea serum nakong ea ho sibolloa ho sebelisa mohlala oa binary logistic regression ka ho netefatsoa ka makhetlo a leshome le ho lekola ts'ebetso e boletsoeng esale pele litsing tsa netefatso tsa kahare le kantle.b Lipalo-palo tse netefalitsoeng tse fapaneng tsa mohlala oa regression oa LASSO bakeng sa khetho ea metabolic biomarker.Linomoro tse fanoeng ka holimo li emela palo e tloaelehileng ea li-biomarker tse khethiloeng ho λ.Mohala o matheba a mafubelu o emela kakaretso ea boleng ba AUC ho lambda e tsamaellanang.Liphoso tsa 'mala o moputsoa li emela bonyane le boholo ba boleng ba AUC.Mohala o nang le matheba o bontša mohlala o motle ka ho fetisisa ka li-biomarker tse 27 tse khethiloeng.AUC, sebaka se tlas'a sebopeho sa moamoheli (ROC) curve.c Mena liphetoho tsa metabolite tse 27 tse khethiloeng sehlopheng sa LA ha li bapisoa le sehlopha sa BN se sehlopheng sa sibollo.Kholomo e khubelu - ts'ebetso.Kholomo e putsoa ke ho fokotseha.d-f Receiver operating character (ROC) curves e bonts'ang matla a mohlala o khethollang ho latela metsoako ea 27 metabolite ho sibolloa, ka hare, le ka ntle ho netefatsa lihlopha.Lintlha tsa mohloli li fanoa ka mokhoa oa lifaele tsa data tsa mohloli.
Moetso oa ho bolela esale pele o entsoe ho ipapisitsoe le li-coefficients tsa boima ba li-metabolite tsena tse 27 (Letlapa la Tlatsetso 3).Tlhahlobo ea ROC e thehiloeng ho li-metabolite tsena tsa 27 e fane ka sebaka se tlas'a boleng ba curve (AUC) ea 0.933, kutloisiso ea sehlopha sa ho sibolla e ne e le 0.868, 'me e khethehileng e ne e le 0.859 (setšoantšo sa 2d).Ho sa le joalo, har'a li-metabolite tse fapaneng tse 38 tse boletsoeng lipakeng tsa LA le HC, sete ea metabolite e 16 e fihletse AUC ea 0.902 ka kutloisiso ea 0.801 le ho khetheha ha 0.856 ho khetholla LA ho HC (Figure Tlatsetso 6a-c).Litekanyetso tsa AUC tse ipapisitseng le litekanyo tse fapaneng tsa phetoho ea foldara bakeng sa metabolite e fapaneng le tsona li ile tsa bapisoa.Re fumane hore mofuta oa lihlopha o sebetsa hantle ka ho fetisisa ho khetholla pakeng tsa LA le BN (HC) ha boemo ba phetoho ea fold e behiloe ho 1.2 khahlanong le 1.5 kapa 2.0 (Figure Supplementary Figure 7a,b).Mokhoa oa ho arola, o thehiloeng ho lihlopha tse 27 tsa metabolite, o ile oa tiisoa hape ka lihlopha tsa ka hare le tsa ka ntle.AUC e ne e le 0.915 (sensitivity 0.867, specificity 0.811) bakeng sa ho netefatsa ka hare le 0.945 (sensitivity 0.810, specificity 0.979) bakeng sa ho netefatsa ka ntle (setšoantšo sa 2e, f).Ho hlahloba katleho ea li-interlaboratory, lisampole tse 40 tse tsoang sehlopheng sa ka ntle li ile tsa hlahlobjoa ka laboratoring e ka ntle joalokaha ho hlalositsoe karolong ea Mekhoa.Ho nepahala ha lihlopha ho fihletse AUC ea 0.925 (Setšoantšo sa Tlatsetso 8).Hobane lung squamous cell carcinoma (LUSC) ke mofuta oa bobeli o atileng haholo oa mofets'e o seng o monyane oa lisele tsa matšoafo (NSCLC) kamora lung adenocarcinoma (LUAD), re boetse re lekile ts'ebeliso e netefalitsoeng ea profil ea metabolic.BN le linyeoe tse 16 tsa LUSC.AUC ea khethollo pakeng tsa LUSC le BN e ne e le 0.776 (Setšoantšo sa Tlatsetso 9), e bontšang bokhoni bo fokolang ha bo bapisoa le khethollo pakeng tsa LUAD le BN.
Liphuputso li bontšitse hore boholo ba li-nodule litšoantšong tsa CT bo amana hantle le monyetla oa ho ba le bothata 'me e ntse e le khetho e kholo ea phekolo ea li-nodule25,26,27.Tlhahlobo ea lintlha tse tsoang sehlopheng se seholo sa thuto ea ho hlahloba NELSON e bontšitse hore kotsi ea ho ba le bothata ba batho ba nang le li-node <5 mm e bile e tšoana le ea lihlooho tse se nang li-node 28.Ka hona, boholo bo fokolang bo hlokang tlhokomelo ea kamehla ea CT ke 5 mm, joalokaha e khothalelitsoe ke British Thoracic Society (BTS), le 6 mm, joalokaha e khothalelitsoe ke Fleischner Society 29.Leha ho le joalo, li-nodule tse kholo ho feta 6 mm le tse se nang litšobotsi tse hlakileng tse nang le bokooa, tse bitsoang indeterminate pulmonary nodules (IPN), e ntse e le phephetso e kholo ho hlahloba le ho laola mekhoa ea bongaka30,31.Ka mor'a moo re ile ra hlahloba hore na boholo ba li-nodule bo susumelitse li-signature tsa metabolomic ho sebelisa lisampole tse kopantsoeng ho tsoa ho li-cohorts tsa ho sibolloa le ho netefatsa ka hare.Re tsepamisitse maikutlo ho li-biomarker tse netefalitsoeng tsa 27, re qalile ho bapisa lintlha tsa PCA tsa HC le BN sub-6 mm metabolomes.Re fumane hore boholo ba lintlha tsa lintlha tsa HC le BN li ne li kopana, ho bontša hore litekanyetso tsa metabolite tsa serum li ne li tšoana le lihlopha ka bobeli (setšoantšo sa 3a).Limmapa tsa likarolo ho pholletsa le mefuta e fapaneng ea boholo li ile tsa lula li bolokiloe BN le LA (setšoantšo sa 3b, c), athe karohano e ile ea hlokomeloa pakeng tsa li-nodule tse kotsi le tse nang le bokooa sebakeng sa 6-20 mm (setšoantšo sa 3d).Sehlopha sena se ne se e-na le AUC ea 0.927, e ikhethang ea 0.868, le kutloisiso ea 0.820 bakeng sa ho bolela esale pele ho senyeha ha li-nodule tse ka bang 6 ho 20 mm (setšoantšo sa 3e, f).Liphetho tsa rona li bonts'a hore sehlopha se ka nka liphetoho tsa metabolic tse bakoang ke phetoho e mpe ea pele, ho sa tsotelehe boholo ba maqhutsu.
ad Papiso ea li-profiles tsa PCA lipakeng tsa lihlopha tse boletsoeng ho ipapisitsoe le sehlopha sa metabolic sa 27 metabolites.CC le BN <6 limilimithara.b BN <6 limilimithara vs BN 6–20 limilimithara.ho LA 6-20 mm khahlanong le LA 20-30 mm.g BN 6-20 limilimithara le LA 6-20 limilimithara.GC, n = 174;BN <6 limilimithara, n = 153;BN 6-20 limilimithara, n = 91;LA 6-20 limilimithara, n = 89;LA 20–30 mm, n = 77. e Receiver operating character (ROC) curve e bontšang ts'ebetso e khethollang ea mohlala bakeng sa li-nodule 6-20 mm.f Litekanyetso tsa menyetla li ne li baloa ho ipapisitsoe le mokhoa oa ho khutlisa thepa bakeng sa li-nodule tse boholo ba 6-20 mm.Mothalo o matheba o mosehla o emela boleng bo nepahetseng ba ho khaoha (0.455).Linomoro tse kaholimo li emetse liperesente tsa linyeoe tse reriloeng bakeng sa Los Angeles.Sebelisa teko ea t ea Moithuti e nang le mehatla e 'meli.PCA, tlhahlobo ea karolo e ka sehloohong.Sebaka sa AUC se tlas'a lekhalo.Lintlha tsa mohloli li fanoa ka mokhoa oa lifaele tsa data tsa mohloli.
Mehlala e mene (ea lilemo tse 44-61) e nang le boholo bo tšoanang ba li-nodule tsa pulmonary (7-9 mm) e ile ea boela ea khethoa ho bontša ts'ebetso ea mokhoa o reriloeng oa ho bolela lintho tse mpe (setšoantšo sa 4a, b).Ha ho hlahlojoa qalong, Nyeoe ea 1 e hlahisitsoe e le nodule e tiileng e nang le calcification, tšobotsi e amanang le botle, athe Nyeoe ea 2 e hlahisitsoe e le nodule e tiileng ka mokhoa o sa lekanyetsoang e se nang litšobotsi tse hlakileng.Likarolo tse tharo tsa CT scans tse latelang li bontšitse hore linyeoe tsena li ile tsa lula li tsitsitse ka nako ea lilemo tse 4 'me kahoo li ne li nkoa e le li-nodule tse nang le benign (setšoantšo sa 4a).Ha ho bapisoa le tlhahlobo ea bongaka ea li-serial CT scans, tlhahlobo e le 'ngoe ea serum metabolite e nang le mohlala oa hona joale oa classifier ka potlako le ka nepo e khethollang maqhutsu ana a nang le bokooa a ipapisitseng le mathata a ka bang teng (Letlapa la 1).Setšoantšo sa 4b tabeng ea 3 se bontša leqhubu le nang le matšoao a pleural retraction, e atisang ho amahanngoa le malignancy32.Nyeoe ea 4 e hlahisoa e le leqhubu le tiileng le sa tsejoeng le se nang bopaki ba sesosa se kotsi.Linyeoe tsena kaofela li ile tsa boleloa esale pele e le tse mpe ho latela mohlala oa classifier (Letlapa la 1).Tlhahlobo ea adenocarcinoma ea matšoafo e ile ea bontšoa ke tlhahlobo ea histopathological ka mor'a ho buuoa ka matšoafo (Setšoantšo sa 4b).Bakeng sa sete ea netefatso ea kantle, sehlopha sa metabolic se boletse ka nepo linyeoe tse peli tsa maqhutsu a mats'oafo a sa feleng a maholo ho feta 6 mm (Setšoantšo sa Tlatsetso 10).
Litšoantšo tsa CT tsa fensetere ea axial ea matšoafo a linyeoe tse peli tsa li-nodule tse nang le benign.Tabeng ea 1, CT scan ka mor'a lilemo tse 4 e bontšitse nodule e tsitsitseng e tiileng e lekanyang 7 mm ka calcification karolong e ka tlaase e tlaase.Tabeng ea 2, CT scan ka mor'a lilemo tse 5 e ile ea senola leqhubu le tsitsitseng, le nang le karolo e itseng e tiileng le bophara ba 7 mm karolong e ka holimo e ka holimo.b Axial fensetere CT litšoantšo tsa matšoafo le tse tsamaellanang pathological lithuto tsa maemo a mabeli a sethaleng I adenocarcinoma pele matšoafo resection.Nyeoe ea 3 e senotse nodule e bophara ba 8 mm karolong e ka holimo e ka ho le letona e nang le pleural retraction.Taba ea 4 e senotse nodule ea khalase e tiileng ea fatše e boholo ba limilimithara tse 9 karolong e kaholimo ea leqele.Hematoxylin le eosin (H&E) matheba a mats'oafo a tsoileng matsoho (scale bar = 50 μm) e bonts'ang mokhoa oa kholo ea acinar ea lung adenocarcinoma.Metsu e bontša li-nodule tse fumanoeng litšoantšong tsa CT.Litšoantšo tsa H & E ke litšoantšo tse emelang likarolo tse ngata (> 3) tse nyenyane tse hlahlojoang ke setsebi sa mafu.
Ha re kopane, liphetho tsa rona li bonts'a boleng bo ka bang teng ba serum metabolite biomarker tlhahlobong e fapaneng ea maqhutsu a matšoafo, a ka bakang mathata ha ho hlahlojoa tlhahlobo ea CT.
Ho ipapisitsoe le phanele e netefalitsoeng ea phapang ea metabolite, re batlile ho tseba li-correlate tsa biological tsa liphetoho tse kholo tsa metabolic.Tlhahlobo ea ntlafatso ea tsela ea KEGG ke MetaboAnalyst e khethileng 6 e tloaelehileng e fetotsoeng litsela tse ngata pakeng tsa lihlopha tse peli tse fanoeng (LA vs. HC le LA vs. BN, e fetotsoeng p ≤ 0.001, phello> 0.01).Liphetoho tsena li ne li khetholloa ke tšitiso ea metabolism ea pyruvate, tryptophan metabolism, niacin le nicotinamide metabolism, glycolysis, potoloho ea TCA, le purine metabolism (Setšoantšo sa 5a).Ka mor'a moo re ile ra tsoela pele ho etsa metabolomics e lebisitsoeng ho netefatsa liphetoho tse kholo ka ho sebelisa palo e feletseng.Boikemisetso ba metabolite e tloaelehileng litseleng tse atisang ho fetoloa ke triple quadrupole mass spectrometry (QQQ) ho sebelisa litekanyetso tsa 'nete tsa metabolite.Litšobotsi tsa palo ea batho ba sampuli ea sepheo sa thuto ea metabolomics li kenyelelitsoe ho Supplementary Table 4. Ho lumellana le liphello tsa rona tsa lefats'e tsa metabolomics, tlhahlobo ea palo e tiisitse hore hypoxanthine le xanthine, pyruvate, le lactate li eketsehile LA ha li bapisoa le BN le HC (Fig. 5b, c, p <0.05).Leha ho le joalo, ha ho liphapang tse kholo ho metabolite tsena tse fumanoeng lipakeng tsa BN le HC.
Tlhahlobo ea ntlafatso ea tsela ea KEGG ea metabolite e fapaneng haholo sehlopheng sa LA ha e bapisoa le lihlopha tsa BN le HC.Ho ile ha sebelisoa Globaltest e nang le mehatla e 'meli,' me litekanyetso tsa p li ile tsa fetoloa ho sebelisoa mokhoa oa Holm-Bonferroni (o fetotsoe p ≤ 0.001 le boholo ba phello> 0.01).b–d Violin plots e bontšang hypoxanthine, xanthine, lactate, pyruvate, le tryptophan level in serum HC, BN, le LA e khethiloeng ke LC-MS/MS (n = 70 ka sehlopha).Mela e matheba a masoeu le a matšo e bontša bohareng le quartile, ka ho latellana.morero oa Violin o bonts'ang Log2TPM e tloaelehileng (lingoloa ka milione) polelo ea mRNA ea SLC7A5 le QPRT ka lung adenocarcinoma (n = 513) ha e bapisoa le lisele tse tloaelehileng tsa matšoafo (n = 59) lethathamong la LUAD-TCGA.Lebokose le lesoeu le emela mefuta ea li-interquartile, mola o motšo o tšekaletseng bohareng o emela bohareng, 'me mola o otlolohileng o motšo o tlohang lebokoseng o emela 95% ea nako ea ho itšepa (CI).f Pearson correlation plot of SLC7A5 le GAPDH expression in lung adenocarcinoma (n = 513) le lithane tse tloaelehileng tsa matšoafo (n = 59) ho dataset ea TCGA.Sebaka se bohlooho se emela 95% CI.r, Pearson coefficient ea khokahano.g Maemo a tloaelehileng a cellular tryptophan liseleng tsa A549 tse fetisitsoeng ka taolo e sa tobang ea shRNA (NC) le shSLC7A5 (Sh1, Sh2) e khethiloeng ke LC-MS/MS.Ho fanoe ka tlhahlobo ea lipalo-palo ea lisampole tse hlano tse ikemetseng ka baeloji sehlopheng ka seng.h Maemo a liselefouno tsa NADt (kakaretso ea NAD, ho kenyeletsoa NAD+ le NADH) ka liseleng tsa A549 (NC) le SLC7A5 li kokota lisele tsa A549 (Sh1, Sh2).Ho fanoe ka tlhahlobo ea lipalo-palo ea lisampole tse tharo tse ikemetseng tsa baeloji sehlopheng ka seng.i Glycolytic mosebetsi oa lisele tsa A549 pele le ka mor'a ho kokota ha SLC7A5 ho ile ha lekanyetsoa ka tekanyo ea extracellular acidification rate (ECAR) (n = 4 disampole tse ikemetseng tsa biologically ka sehlopha).2-DG, 2-deoxy-D-glucose.Teko ea Moithuti e nang le mehatla e 'meli e sebelisitsoe ho (b–h).Ho (g-i), mekoallo ea liphoso e emela se boleloang ke ± SD, teko e 'ngoe le e' ngoe e entsoe ka makhetlo a mararo ka boithaopo 'me liphello li tšoana.Lintlha tsa mohloli li fanoa ka mokhoa oa lifaele tsa data tsa mohloli.
Ha re nahana ka phello e kholo ea metabolism ea tryptophan e fetotsoeng sehlopheng sa LA, re ile ra boela ra hlahloba maemo a serum tryptophan ho lihlopha tsa HC, BN, le LA re sebelisa QQQ.Re fumane hore serum tryptophan e fokotsehile ho LA ha e bapisoa le HC kapa BN (p <0.001, Figure 5d), e lumellanang le liphuputso tse fetileng tsa hore litekanyetso tsa tryptophan tse potolohang li fokotsehile ho bakuli ba nang le kankere ea matšoafo ho feta ho laola bophelo bo botle ho tsoa sehlopheng sa taolo33,34 ,35.Phuputso e 'ngoe e sebelisang PET/CT tracer 11C-methyl-L-tryptophan e fumane hore nako ea ho boloka lets'oao la tryptophan ka har'a lisele tsa mofetše oa matšoafo e eketsehile haholo ha e bapisoa le liso tse mpe kapa lisele tse tloaelehileng36.Re nahana hore ho fokotseha ha tryptophan ho LA serum ho ka bonts'a ts'ebeliso e sebetsang ea tryptophan ke lisele tsa mofetše oa matšoafo.
Ho boetse hoa tsebahala hore sehlahisoa sa ho qetela sa tsela ea kynurenine ea tryptophan catabolism ke NAD+37,38, e leng karolo ea bohlokoa bakeng sa karabelo ea glyceraldehyde-3-phosphate e nang le 1,3-bisphosphoglycerate ho glycolysis39.Leha lithuto tse fetileng li tsepamisitse maikutlo ho karolo ea tryptophan catabolism taolong ea 'mele ea ho itšireletsa mafung, re ile ra batla ho hlakisa tšebelisano lipakeng tsa tryptophan dysregulation le litsela tsa glycolytic tse hlokometsoeng thutong ea hajoale.Solute transporter family 7 member 5 (SLC7A5) e tsejoa e le tryptophan transporter43,44,45.Quinolinic acid phosphoribosyltransferase (QPRT) ke enzyme e fumanehang ka tlase ho tsela ea kynurenine e fetolang quinolinic acid ho NAMN46.Tlhahlobo ea dataset ea LUAD TCGA e senoletse hore SLC7A5 le QPRT ka bobeli li ne li laoloa haholo ke lisele tsa tumor ha li bapisoa le lisele tse tloaelehileng (Fig. 5e).Keketseho ena e ile ea bonoa ka mekhahlelo ea I le II hammoho le mekhahlelo ea III le IV ea lung adenocarcinoma (Figure ea Tlatsetso 11), e bontšang tšitiso ea pele ea tryptophan metabolism e amanang le tumorigenesis.
Ntle le moo, datha ea LUAD-TCGA e bonts'itse khokahano e ntle lipakeng tsa SLC7A5 le polelo ea GAPDH mRNA lisampole tsa bakuli ba mofets'e (r = 0.45, p = 1.55E-26, Figure 5f).Ka lehlakoreng le leng, ha ho na kamano ea bohlokoa e fumanoeng pakeng tsa liphatsa tsa lefutso tse joalo ka lisele tse tloaelehileng tsa matšoafo (r = 0.25, p = 0.06, Figure 5f).Knockdown ea SLC7A5 (Supplementary Figure 12) liseleng tsa A549 e fokolitse haholo li-tryptophan tsa cellular le NAD (H) (Setšoantšo sa 5g, h), e leng se bakang ts'ebetso ea glycolytic e fokolisitsoeng joalokaha e lekantsoe ka tekanyo ea extracellular acidification rate (ECAR) (Setšoantšo sa 1).5i).Kahoo, ho ipapisitse le liphetoho tsa metabolic ho serum le ho lemoha ha in vitro, re nahana hore tryptophan metabolism e ka hlahisa NAD+ ka tsela ea kynurenine mme ea bapala karolo ea bohlokoa ho khothaletseng glycolysis mofetšeng oa matšoafo.
Liphuputso li bontšitse hore palo e kholo ea li-nodule tsa pulmonary tse sa tsejoeng tse fumanoeng ke LDCT li ka 'na tsa lebisa tlhokahalong ea tlhahlobo e eketsehileng e kang PET-CT, lung biopsy, le ho phekola ka ho feteletseng ka lebaka la ho hlahlojoa ka bohata ha lefu la 31 Joalokaha ho bontšitsoe setšoantšong sa 6. boithuto ba rona bo bonts'itse sehlopha sa metabolite ea serum e nang le boleng ba tlhahlobo e ka ntlafatsang stratification ea kotsi le taolo e latelang ea li-nodule tsa pulmonary tse fumanoeng ke CT.
Li-nodule tsa matšoafo li hlahlojoa ho sebelisoa tekanyo e tlaase ea computed tomography (LDCT) e nang le likarolo tsa litšoantšo tse bontšang lisosa tse mpe kapa tse kotsi.Phello e sa tsitsang ea li-nodule e ka lebisa maetong a ho latella khafetsa, ho kenella ho sa hlokahaleng, le phekolo e feteletseng.Ho kenyelletsoa ha li-classifiers tsa serum metabolic tse nang le boleng ba tlhahlobo ho ka ntlafatsa tlhahlobo ea kotsi le taolo e latelang ea li-nodule tsa pulmonary.PET positron emission tomography.
Lintlha tse tsoang thutong ea NLST ea US le thuto ea NELSON ea Europe li fana ka maikutlo a hore ho hlahloba lihlopha tse kotsing e kholo ka tekanyo e tlaase ea computed tomography (LDCT) ho ka fokotsa lefu la kankere ea matšoafo1,3.Leha ho le joalo, tlhahlobo ea kotsi le taolo e latelang ea kliniki ea palo e kholo ea maqhutsu a pulmonary a fumanoeng ke LDCT e ntse e le phephetso ka ho fetisisa.Sepheo sa mantlha ke ho ntlafatsa likarolo tse nepahetseng tsa liprothokholo tse teng tse thehiloeng ho LDCT ka ho kenyelletsa li-biomarker tse tšepahalang.
Li-biomarker tse ling tsa limolek'hule, joalo ka metabolite ea mali, li fumanoe ka ho bapisa mofetše oa matšoafo le taolo e phetseng hantle15,17.Thutong ea hajoale, re tsepamisitse maikutlo ho ts'ebeliso ea tlhahlobo ea serum metabolomics ho khetholla lipakeng tsa maqhutsu a mabe le a kotsi a pulmonary a fumanoeng ka tšohanyetso ke LDCT.Re bapisitse metabolome ea lefats'e ea serum ea taolo e phetseng hantle (HC), li-nodule tsa benign lung (BN), le lisampole tsa sethala sa I lung adenocarcinoma (LA) re sebelisa tlhahlobo ea UPLC-HRMS.Re fumane hore HC le BN li na le litlaleho tse tšoanang tsa metabolic, athe LA e bontšitse liphetoho tse kholo ha li bapisoa le HC le BN.Re khethile lihlopha tse peli tsa metabolite ea serum e khethollang LA ho HC le BN.
Morero oa hajoale oa ho tsebahatsa o thehiloeng ho LDCT bakeng sa maqhutsu a kotsi le a kotsi o ipapisitse haholo le boholo, letsoalo, sebopeho le sekhahla sa kholo ea maqhutsu ka nako30.Liphuputso tse fetileng li bontšitse hore boholo ba maqhutsu bo amana haufi-ufi le monyetla oa ho ba le mofetše oa matšoafo.Esita le ho bakuli ba kotsing e kholo, kotsi ea ho ba le bokooa ho nodes <6 mm ke <1%.Kotsi ea lefu la maqhutsu a boholo ba 6 ho isa ho 20 mm e tloha ho 8% ho isa ho 64%30.Ka hona, Mokhatlo oa Fleischner o khothalletsa ho fokotsa bophara ba 6 mm bakeng sa ho latela mokhoa o tloaelehileng oa CT.29 Leha ho le joalo, tlhahlobo ea kotsi le tsamaiso ea li-nodule tse sa feleng tsa pulmonary (IPN) tse kholo ho feta 6 mm ha lia etsoa ka ho lekaneng 31 .Tsamaiso ea hona joale ea lefu la pelo ea congenital hangata e itšetlehile ka ho leta ka hloko ka tlhahlobo ea CT khafetsa.
Ho latela metabolome e netefalitsoeng, re bonts'itse ka lekhetlo la pele ho kopana ha li-signature tsa metabolomic lipakeng tsa batho ba phetseng hantle le li-nodule tse bobebe <6 mm.Ho tšoana ha likokoana-hloko ho lumellana le liphuputso tse fetileng tsa CT hore kotsi ea ho senya li-nodules <6 mm e tlaase joaloka lihlooho tse se nang li-node.30 Re lokela ho hlokomela hore liphetho tsa rona li boetse li bontša hore li-nodule tse nang le benign <6 mm le ≥6 mm li na le phahameng. ho tšoana ho litlaleho tsa metabolomic, ho fana ka maikutlo a hore tlhaloso e sebetsang ea benign etiology e lumellana ho sa tsotellehe boholo ba nodule.Ka hona, liphanele tsa morao-rao tsa ho hlahloba serum metabolite li ka fana ka tlhahlobo e le 'ngoe e le teko ea molao ha li-nodule li qala ho fumanoa ho CT 'me li ka fokotsa ho hlahlojoa ha serial.Ka nako e ts'oanang, sehlopha se le seng sa li-biomarker tsa metabolism li ile tsa khetholla li-nodule tse kotsi ≥6 limilimithara ka boholo ho tloha ho li-nodule tse nang le bokooa 'me li fana ka likhakanyo tse nepahetseng bakeng sa li-IPN tsa boholo bo tšoanang le likarolo tse sa hlakang tsa morphological litšoantšong tsa CT.Sehlopha sena sa serum metabolism se sebelitse hantle ha se bolela esale pele bobe ba maqhutsu ≥6 mm ka AUC ea 0.927.Ha re kopane, liphetho tsa rona li bonts'a hore li-signature tse ikhethang tsa serum metabolomic li ka bonts'a ka ho hlaka liphetoho tsa metabolic tse bakiloeng ke tumor mme li na le boleng bo ka bang teng e le lits'oants'o tsa kotsi, ntle le boholo ba maqhutsu.
Haholo-holo, lung adenocarcinoma (LUAD) le squamous cell carcinoma (LUSC) ke mefuta ea mantlha ea mofetše o seng o monyane oa lisele tsa matšoafo (NSCLC).Ka lebaka la hore LUSC e amahanngoa haholo le ts'ebeliso ea koae47 mme LUAD ke nalane e atileng haholo ea li-nodule tsa matšoafo tse bonoang tlhahlobong ea CT48, mofuta oa rona oa sehlopha o hahiloe ka ho khetheha bakeng sa disampole tsa sethala sa I adenocarcinoma.Wang le basebetsi-'moho le bona ba tsepamisitse maikutlo ho LUAD mme ba tsebahatsa li-signature tse robong tsa lipid tse sebelisang lipidomics ho khetholla mofetše oa matšoafo oa pele ho batho ba phetseng hantle17.Re lekile mofuta oa hajoale oa sehlopha sa linyeoe maemong a 16 a sethala sa I LUSC le li-nodule tse 74 mme ra bona bonnete bo tlase ba LUSC (AUC 0.776), re fana ka maikutlo a hore LUAD le LUSC li kanna tsa ba le li-signature tsa tsona tsa metabolomic.Ehlile, LUAD le LUSC li bonts'itsoe hore li fapane ho etiology, tšimoloho ea tlhaho le liphatsa tsa lefutso49.Ka hona, mefuta e meng ea histology e lokela ho kenyelletsoa mefuteng ea koetliso bakeng sa ho lemoha mofetše oa matšoafo o thehiloeng ho baahi mananeong a tlhahlobo.
Mona, re khethile litsela tse tšeletseng tse fetotsoeng hangata ho lung adenocarcinoma ha li bapisoa le li-control tse phetseng hantle le li-nodule tse bobebe.Xanthine le hypoxanthine ke metabolites tse tloaelehileng tsa tsela ea metabolic ea purine.Tumellanong le liphello tsa rona, li-intermediate tse amanang le metabolism ea purine li ile tsa eketseha haholo ka serum kapa lisele tsa bakuli ba nang le lung adenocarcinoma ha li bapisoa le ho laola bophelo bo botle kapa bakuli ba sethaleng sa preinvasive15,50.Maemo a phahameng a serum xanthine le hypoxanthine a ka bonts'a anabolism e hlokoang ke lisele tsa mofetše tse ntseng li ata ka potlako.Ho senyeha ha metabolism ea tsoekere ke letšoao le tsebahalang la metabolism ea mofetše51.Mona, re bone keketseho e kholo ea pyruvate le lactate sehlopheng sa LA ha se bapisoa le sehlopha sa HC le BN, se tsamaellanang le litlaleho tse fetileng tsa ho se tloaelehe ha tsela ea glycolytic profilaneng ea serum metabolome ea bakuli bao e seng ba mofets'e oa mofets'e oa matšoafo (NSCLC) le ditaolo tse phetseng hantle.liphetho li lumellana52,53.
Habohlokoa, re bone khokahano e fapaneng lipakeng tsa pyruvate le tryptophan metabolism serum ea lung adenocarcinomas.Maemo a serum tryptophan a fokotsehile sehlopheng sa LA ha a bapisoa le sehlopha sa HC kapa BN.Hoa thahasellisa hore phuputso e fetileng ea boholo-holo e sebelisang sehlopha se neng se tla ba teng e fumane hore litekanyetso tse tlaase tsa ho potoloha ha tryptophan li ne li amahanngoa le kotsi e eketsehileng ea kankere ea matšoafo 54.Tryptophan ke amino acid ea bohlokoa eo re e fumanang lijong.Re etsa qeto ea hore serum tryptophan depletion ka lung adenocarcinoma e ka 'na ea bontša ho fokotseha ka potlako ha metabolite ena.Hoa tsebahala hore sehlahisoa sa ho qetela sa tryptophan catabolism ka tsela ea kynurenine ke mohloli oa de novo NAD + synthesis.Hobane NAD+ e hlahisoa haholo-holo ka tsela ea pholoso, bohlokoa ba NAD+ ho tryptophan metabolism ho bophelo bo botle le mafu bo ntse bo lokela ho boleloa46.Tlhahlobo ea rona ea database ea TCGA e bontšitse hore polelo ea tryptophan transporter solute transporter 7A5 (SLC7A5) e eketsehile haholo ho lung adenocarcinoma ha e bapisoa le taolo e tloaelehileng mme e ne e amana hantle le polelo ea enzyme ea glycolytic GAPDH.Lithuto tse fetileng li shebile haholo karolo ea tryptophan catabolism ho hatella karabelo ea 'mele ea ho itšireletsa mafung ea antitumor40,41,42.Mona re bonts'a hore ho thibela ho ts'oaroa ha tryptophan ka ho kokota ha SLC7A5 liseleng tsa mofetše oa matšoafo ho fella ka phokotso e latelang ea maemo a cellular a NAD le ho fokotseha ho hong ha ts'ebetso ea glycolytic.Ka bokhutšoanyane, thuto ea rona e fana ka motheo oa likokoana-hloko bakeng sa liphetoho tsa metabolism ea serum e amanang le phetoho e mpe ea lung adenocarcinoma.
Liphetoho tsa EGFR ke liphetoho tse atileng haholo ho bakuli ba NSCLC.Phuputsong ea rona, re fumane hore bakuli ba nang le phetoho ea EGFR (n = 41) ba ne ba e-na le litlaleho tse akaretsang tsa metabolomic tse tšoanang le bakuli ba nang le mofuta oa hlaha oa EGFR (n = 31), le hoja re fumane ho fokotseha ha maemo a serum ho bakuli ba bang ba EGFR ba fetohileng ho bakuli ba acylcarnitine.Mosebetsi o thehiloeng oa li-acylcarnitine ke ho tsamaisa lihlopha tsa acyl ho tloha cytoplasm ho ea matrix ea mitochondrial, e lebisang ho oxidation ea mafura a mafura ho hlahisa matla a 55.Tumellanong le seo re se fumaneng, phuputso ea morao-rao e boetse e fumane litlaleho tse tšoanang tsa metabolome pakeng tsa EGFR mutant le EGFR mofuta oa hlaha oa lihlahala ka ho hlahloba metabolome ea lefats'e ea 102 lung adenocarcinoma tissue sampuli50.Hoa thahasellisa hore litaba tsa acylcarnitine li ile tsa boela tsa fumanoa sehlopheng sa EGFR mutant.Ka hona, hore na liphetoho tsa maemo a acylcarnitine li bonahatsa liphetoho tsa metabolic tse bakoang ke EGFR le litsela tse ka tlaase tsa limolek'hule li ka tšoanela ho ithuta ho eketsehileng.
Qetellong, boithuto ba rona bo theha serum metabolic classifier bakeng sa tlhahlobo e fapaneng ea li-nodule tsa pulmonary mme e fana ka tlhahiso ea phallo ea mosebetsi e ka ntlafatsang tlhahlobo ea kotsi le ho tsamaisa taolo ea tleliniki e ipapisitseng le tlhahlobo ea CT scan.
Boithuto bona bo amohetsoe ke Komiti ea Boitšoaro ea Sepetlele sa Kankere sa Univesithi ea Sun Yat-sen, Sepetlele sa Pele se Kopanetsoeng sa Univesithi ea Sun Yat-sen, le Komiti ea Boitšoaro ea Sepetlele sa Kankere sa Univesithi ea Zhengzhou.Lihlopheng tsa ho sibolloa le ho netefatsa ka hare, li-sera tse 174 tse tsoang ho batho ba phetseng hantle le li-sera tse 244 tse tsoang ho li-nodule tse nang le bokooa li ile tsa bokelloa ho batho ba neng ba etsoa litlhahlobo tsa selemo le selemo Lefapheng la Taolo le Thibelo ea Kankere, Setsing sa Kankere sa Univesithi ea Sun Yat-sen, le li-nodule tse 166 tse kotsi.serum.Stage I lung adenocarcinomas e ile ea bokelloa Setsing sa Kankere sa Univesithi ea Sun Yat-sen.Sehlopheng sa ho netefatsa ka ntle, ho ne ho e-na le linyeoe tse 48 tsa li-nodule tse nang le benign, linyeoe tse 39 tsa sethaleng sa I lung adenocarcinoma ho tloha Sepetleleng sa Pele se Kopanetsoeng sa Univesithi ea Sun Yat-sen, le linyeoe tse 24 tsa sethaleng sa I lung adenocarcinoma ho tsoa Sepetleleng sa Kankere sa Zhengzhou.Setsi sa Kankere sa Univesithi ea Sun Yat-sen le sona se ile sa bokella linyeoe tse 16 tsa mofets'e oa mofets'e oa mofets'e oa sele ea squamous ho lekola bokhoni ba tlhahlobo ea sehlopha sa metabolic se thehiloeng (litšobotsi tsa mokuli li bonts'itsoe ho Letlapa la Tlatsetso la 5).Mehlala ho tsoa ho sehlopha sa phumano le sehlopha sa netefatso ea ka hare e ile ea bokelloa lipakeng tsa Pherekhong 2018 le Mots'eanong 2020. Mehlala ea sehlopha sa netefatso ea kantle e ile ea bokelloa lipakeng tsa Phato 2021 le Mphalane 2022. Ho fokotsa leeme la bong, palo e ka bang lekanang ea linyeoe tsa banna le basali li abetsoe e 'ngoe le e 'ngoe. sehlopha.Sehlopha sa Discovery and Internal Review Team.Bong ba morupeluoa bo ile ba khethoa ho ipapisitsoe le boitlamo ba bona.Tumello e nang le tsebo e ile ea fumanoa ho barupeluoa bohle 'me ha ho matšeliso a fanoeng.Lihlooho tse nang le li-nodule tse nang le benign e ne e le tse nang le lintlha tse tsitsitseng tsa CT scan ho 2 ho lilemo tse 5 nakong ea tlhahlobo, ntle le nyeoe ea 1 e tsoang ho sampuli ea ka ntle ea netefatso, e ileng ea bokelloa pele ho ts'ebetso le ho fumanoa ke histopathology.Ntle le bronchitis e sa foleng.Linyeoe tsa adenocarcinoma tsa matšoafo li ile tsa bokelloa pele ho ts'ebetso ea matšoafo 'me tsa tiisoa ke tlhahlobo ea pathological.Lisampole tsa mali tse itima lijo li ile tsa bokelloa ka har'a li-tubes tsa karohano ea serum ntle le li-anticoagulants.Mehlala ea mali e ile ea koaheloa ka hora e le 'ngoe ka mocheso oa kamore' me ea centrifuged ho 2851 × g ka metsotso e 10 ho 4 ° C ho bokella serum supernatant.Li-aliquots tsa serum li ne li hoamisitsoe ho -80°C ho fihlela ho ntšoa ha metabolite.Lefapha la Thibelo ea Kankere le Tlhahlobo ea Bongaka la Setsi sa Kankere sa Univesithi ea Sun Yat-sen le ile la bokella letamo la serum ho tsoa ho bafani ba 100 ba phetseng hantle, ho kenyelletsa le palo e lekanang ea banna le basali ba lilemo tse 40 ho isa ho tse 55.Melamo e lekanang ea sampole e 'ngoe le e 'ngoe ea bafani e ile ea tsoakoa, letamo le hlahisitsoeng le ile la qotsoa 'me la bolokoa ho -80 ° C.Motsoako oa serum o ne o sebelisoa e le lisebelisoa bakeng sa taolo ea boleng le maemo a data.
Serum ea litšupiso le li-sampuli tsa liteko li ile tsa qhibiliha 'me metabolite e ntšoa ka mokhoa o kopantsoeng oa ho ntša (MTBE / methanol / metsi) 56.Ka bokhutšoanyane, 50 μl ea serum e kopantsoe le 225 μl ea methanol e batang ka leqhoa le 750 μl ea leqhoa le batang la methyl tert-butyl ether (MTBE).Hlohlelletsa motsoako 'me u e kenye holim'a leqhoa bakeng sa 1 hora.Joale lisampole li ne li tsoakoa 'me vortex e kopantsoe le 188 μl ea metsi a grade ea MS a nang le litekanyetso tsa ka hare (13C-lactate, 13C3-pyruvate, 13C-methionine, le 13C6-isoleucine, e rekiloeng ho Cambridge Isotope Laboratories).Ka nako eo motsoako o ne o le centrifuged ho 15,000 × g bakeng sa 10 min ho 4 ° C, 'me karolo e tlaase e fetisetsoa ka li-tubes tse peli (125 μL ka' ngoe) bakeng sa tlhahlobo ea LC-MS ka mekhoa e metle le e mpe.Qetellong, sampole e ile ea fetoha mouoane hore e omelle ka har'a sekontiri sa vacuum sa lebelo le holimo.
Li-metabolite tse omisitsoeng li ile tsa tsosolosoa ka 120 μl ea 80% ea acetonitrile, ea vortexed bakeng sa metsotso ea 5, 'me ea centrifuged ho 15,000 × g bakeng sa 10 min ho 4 ° C.Li-supernatants li ile tsa fetisetsoa ka har'a linkho tsa khalase ea amber tse nang le li-microinserts bakeng sa lithuto tsa metabolomics.Tlhahlobo e sa lebelloang ea metabolomics sethaleng sa ultra-performance liquid chromatography-high-resolution mass spectrometry (UPLC-HRMS) sethaleng.Li-metabolites li ne li arohane ka ho sebelisa tsamaiso ea Dionex Ultimate 3000 UPLC le kholumo ea ACQUITY BEH Amide (2.1 × 100 mm, 1.7 μm, Metsi).Ka mokhoa o motle oa ion, mekhahlelo ea mobile e ne e le 95% (A) le 50% acetonitrile (B), e 'ngoe le e' ngoe e na le 10 mmol / L ammonium acetate le 0.1% formic acid.Ka mokhoa o mobe, mekhahlelo ea mobile A le B e ne e e-na le 95% le 50% acetonitrile, ka ho latellana, mekhahlelo ka bobeli e na le 10 mmol / L ammonium acetate, pH = 9. Lenaneo la gradient le ne le le tjena: 0-0.5 min, 2% B;0.5–12 mets, 2–50% B;12–14 mets, 50–98% B;14–16 mets, 98% B;16–16.1.mets, 98 –2% B;16.1-20 min, 2% B. Kholomo e ne e bolokiloe ho 40 ° C le sampuli ho 10 ° C ho autosampler.Sekhahla sa phallo e ne e le 0.3 ml / min, molumo oa ente e ne e le 3 μl.Q-Exactive Orbitrap mass spectrometer (Thermo Fisher Scientific) e nang le mohloli oa electrospray ionization (ESI) e ne e sebelisoa ka mokhoa o feletseng oa ho hlahloba 'me o kopantsoe le mokhoa oa ho beha leihlo ddMS2 ho bokella lintlha tse ngata.Litekanyetso tsa MS li ne li behiloe ka tsela e latelang: motlakase oa spray +3.8 kV/- 3.2 kV, mocheso oa capillary 320 ° C, khase e sireletsang 40 arb, khase e thusang 10 arb, probe heater mocheso 350 ° C, scanning range 70-1050 m/h, qeto.70 000. Lintlha li ile tsa fumanoa ho sebelisoa Xcalibur 4.1 (Thermo Fisher Scientific).
Ho lekola boleng ba data, lisampole tsa taolo ea boleng bo kopaneng (QC) li entsoe ka ho tlosa li-aliquots tsa 10 μL tsa supernatant ho sampole ka 'ngoe.Liente tse tšeletseng tsa litekanyetso tsa boleng li ile tsa hlahlojoa qalong ea tatellano ea tlhahlobo ho lekola botsitso ba sistimi ea UPLC-MS.Joale lisampole tsa taolo ea boleng li kenngoa ka har'a beche nako le nako.Kaofela lihlopha tse 11 tsa lisampole tsa serum thutong ena li ile tsa hlahlobjoa ke LC-MS.Li-aliquots tsa motsoako oa letamo la serum ho tsoa ho bafani ba 100 ba phetseng hantle li ile tsa sebelisoa e le litšupiso ka lihlopha tse fapaneng ho lekola ts'ebetso ea ho ntšoa le ho ikamahanya le litlamorao tsa batch-to-batch.Tlhahlobo e sa lebelloang ea metabolomics ea sehlopha sa ho sibolla, sehlopha sa netefatso ea kahare, le sehlopha sa netefatso sa kantle se ile sa etsoa Setsing sa Metabolomics sa Univesithi ea Sun Yat-sen.Laeborari e kantle ea Setsi sa Tlhahlobo le Teko sa Univesithi ea Guangdong le eona e ile ea hlahlobisisa lisampole tse 40 ho tsoa sehlopheng sa kantle ho leka ts'ebetso ea mofuta oa lihlopha.
Ka mor'a ho ntšoa le ho tsosolosoa, palo e feletseng ea metabolite ea serum e ile ea lekanyetsoa ho sebelisoa ultra-high performance liquid chromatography-tandem mass spectrometry (Agilent 6495 triple quadrupole) e nang le mohloli oa electrospray ionization (ESI) ka mokhoa oa multiple reaction monitoring (MRM).Kholomo ea ACQUITY BEH Amide (2.1 × 100 mm, 1.7 μm, Metsi) e ne e sebelisetsoa ho arola metabolites.Karolo ea selefouno e ne e e-na le 90% (A) le 5% ea acetonitrile (B) e nang le 10 mmol / L ammonium acetate le 0.1% ea tharollo ea ammonia.Lenaneo la gradient le ne le le tjena: 0–1.5 min, 0% B;1.5–6.5 mets, 0–15% B;6.5-8 mets, 15% B;8–8.5 mets, 15%–0% B;8.5–11.5 mets, 0%B.Kholomo e ne e hlokometsoe ho 40 °C le sampole ho 10 °C ho lisampler ea likoloi.Sekhahla sa phallo e ne e le 0.3 mL / min mme molumo oa ente e ne e le 1 μL.Litekanyetso tsa MS li ne li behiloe ka tsela e latelang: matla a capillary ± 3.5 kV, khatello ea nebulizer 35 psi, khase ea sheath e phallang 12 L / min, mocheso oa khase ea sheath 350 ° C, mocheso oa khase o omisang 250 ° C, le ho omisa khase ea 14 l / min.Liphetoho tsa MRM tsa tryptophan, pyruvate, lactate, hypoxanthine le xanthine e ne e le 205.0–187.9, 87.0–43.4, 89.0–43.3, 135.0–92.3 le 151.0–107.9 ka ho latellana.Lintlha li ile tsa bokelloa ho sebelisoa Mass Hunter B.07.00 (Agilent Technologies).Bakeng sa disampole tsa serum, tryptophan, pyruvate, lactate, hypoxanthine, le xanthine li ile tsa lekanyetsoa ho sebelisoa li-curve tsa calibration tsa tharollo e tloaelehileng ea motsoako.Bakeng sa lisampole tsa lisele, litaba tsa tryptophan li ne li tloaeleha ho ea ho maemo a ka hare le boima ba protheine ea sele.
Ho ntšoa ha tlhōrō (m/z le nako ea ho boloka (RT)) ho entsoe ho sebelisoa Compound Discovery 3.1 le TraceFinder 4.0 (Thermo Fisher Scientific).Ho felisa liphapang tse ka bang teng lipakeng tsa lihlopha, tlhoro ea mofuta o mong le o mong oa sampole ea teko e ile ea aroloa ke tlhoro e ikhethang ea thepa ea litšupiso ho tsoa sehlopheng se le seng ho fumana bongata bo lekanang.Liphapang tse amanang le maemo a ka hare pele le ka mor'a maemo li bonts'oa ho Tlatsetso Lethathamo la 6. Liphapang pakeng tsa lihlopha tsena tse peli li ne li khetholloa ka tekanyo ea bohata ea ho sibolloa (FDR<0.05, teko ea boemo bo saenneng ea Wilcoxon) le ho fetoha ha menahano (> 1.2 kapa <0.83).Lintlha tse tala tsa MS tsa likarolo tse ntšitsoeng le lintlha tsa litšupiso tsa MS tse lokiselitsoeng serum li bonts'oa ho Supplementary Data 1 le Supplementary Data 2, ka ho latellana.Tlhaloso ea tlhōrō e ile ea etsoa ho latela litekanyetso tse 'nè tse hlalositsoeng tsa boitsebiso, ho akarelletsa le metabolite e khethiloeng, metsoako ea putatively annotated, putatively characterized compound classes, le metsoako e sa tsejoeng 22.Ho ipapisitsoe le lipatlisiso tsa database ho Compound Discovery 3.1 (mzCloud, HMDB, Chemspider), metsoako ea likokoana-hloko e nang le MS/MS e lumellanang le litekanyetso tse tiisitsoeng kapa litlhaloso tse nepahetseng tsa papali ho mzCloud (lintlha> 85) kapa Chemspider qetellong li ile tsa khethoa e le li-intermediate pakeng tsa metabolome e fapaneng.Litlhaloso tse phahameng tsa karolo e 'ngoe le e' ngoe li kenyelelitsoe ho Supplementary Data 3. MetaboAnalyst 5.0 e sebelisitsoe bakeng sa tlhahlobo e sa fetoheng ea bongata ba metabolite e tloaelehileng.MetaboAnalyst 5.0 e boetse e hlahlobile tlhahlobo ea ntlafatso ea tsela ea KEGG e thehiloeng ho metabolite e fapaneng haholo.Principal component analysis (PCA) le partial least squares discriminant analysis (PLS-DA) li ile tsa hlahlojoa ho sebelisoa ropls software package (v.1.26.4) le stack normalization le autoscaling.The optimal metabolite biomarker model bakeng sa ho bolela esale pele lefu la nodule le ile la hlahisoa ho sebelisoa binary logistic regression le bonyane ka ho feletseng shrinkage le khetho opareitara (LASSO, R package v.4.1-3).Ts'ebetso ea mohlala oa khethollo ho lihlopha tsa ho lemoha le ho netefatsa e ne e khetholloa ka ho lekanya AUC ho latela tlhahlobo ea ROC ho latela sephutheloana sa pROC (v.1.18.0.).Monyetla o motle oa ho khaola o ile oa fumanoa ho ipapisitsoe le index e phahameng ea Youden ea mohlala (sensitivity + specificity - 1).lisampole tse nang le boleng bo tlase kapa tse kholo ho feta moeli li tla boleloa esale pele e le li-nodule tse bobebe le lung adenocarcinoma, ka ho latellana.
Lisele tsa A549 (#CCL-185, Pokello ea Setso sa Mofuta oa Amerika) li ne li lengoa ka mokhoa oa F-12K o nang le 10% FBS.Li-hairpin tse khutšoane tsa RNA (shRNA) tse lebisitseng ho SLC7A5 le taolo e sa lebelloang (NC) li kentsoe ho lentiviral vector pLKO.1-puro.Tatelano ea antisense ea shSLC7A5 ke e latelang: Sh1 (5′-GGAGAAACCTGATGAACAGTT-3′), Sh2 (5′-GCCGTGGACTTCGGGAACTAT-3′).Li-antibodies ho SLC7A5 (#5347) le tubulin (#2148) li ile tsa rekoa ho Cell Signaling Technology.Li-antibodies ho SLC7A5 le tubulin li ne li sebelisoa ka ho hlapolloa ha 1: 1000 bakeng sa tlhahlobo ea blot ea Bophirimela.
Teko ea Seahorse XF Glycolytic Stress Test e lekanya maemo a extracellular acidification (ECAR).Tekong, tsoekere, oligomycin A, le 2-DG li ile tsa tsamaisoa ka tatellano ho leka bokhoni ba cellular glycolytic joalo ka ha bo lekantsoe ke ECAR.
Lisele tsa A549 tse fetisitsoeng ka taolo e sa lebelloang (NC) le shSLC7A5 (Sh1, Sh2) li ne li pentiloe ka bosiu bo le bong ka lijana tsa bophara ba 10 cm.Li-metabolite tsa lisele li ile tsa ntšoa ka 1 ml ea leqhoa le batang la 80% la methanol ea metsi.Lisele tse ka har'a tharollo ea methanol li ile tsa hlajoa, tsa bokelloa ka har'a tube e ncha, 'me tsa kenngoa bohareng ho 15,000 × g bakeng sa metsotso e 15 ho 4 ° C.Bokella 800 µl ea matla a phahametseng tlhaho 'me u omelle u sebelisa sekontiri sa vacuum sa lebelo le holimo.Li-pellets tse omisitsoeng tsa metabolite li ile tsa hlahlojoa bakeng sa litekanyetso tsa tryptophan ho sebelisa LC-MS/MS joalokaha ho hlalositsoe ka holimo.Maemo a Cellular NAD(H) liseleng tsa A549 (NC le shSLC7A5) a lekantsoe ho sebelisoa NAD+/NADH colorimetric kit (#K337, BioVision) ho latela litaelo tsa moetsi.Maemo a protheine a lekantsoe bakeng sa sampole ka 'ngoe ho etsa hore palo ea metabolite e tloaelehe.
Ha ho mekhoa ea lipalo-palo e ileng ea sebelisoa ho fumana boholo ba sampole esale pele.Lithuto tsa nakong e fetileng tsa metabolomics tse reretsoeng ho sibolloa ha biomarker15,18 li nkuoe e le litekanyetso tsa ho fumana boholo, 'me ha li bapisoa le litlaleho tsena, sampole ea rona e ne e lekane.Ha ho lisampole tse qheletsoeng ka thoko ho sehlopha sa boithuto.Li-sampuli tsa serum li ne li abeloa ka mokhoa o sa reroang ho sehlopha sa ho sibolla (linyeoe tsa 306, 74.6%) le sehlopha sa ho netefatsa ka hare (maemo a 104, 25.4%) bakeng sa lithuto tsa metabolomics tse sa lebelloang.Re boetse re khethile ka mokhoa o sa reroang linyeoe tse 70 ho tsoa sehlopheng ka seng ho tsoa ho sibollo e behiloeng bakeng sa lithuto tse lebisitsoeng tsa metabolomics.Bafuputsi ba ile ba foufatsoa ke mosebetsi oa sehlopha nakong ea ho bokella le ho hlahloba lintlha tsa LC-MS.Litlhahlobo tsa lipalo-palo tsa data ea metabolomics le liteko tsa lisele li hlalositsoe likarolong tse fapaneng tsa Liphetho, Litlaleho tsa Litšoantšo, le Mekhoa.Tekanyetso ea ts'ebetso ea cellular tryptophan, NADT, le glycolytic e entsoe ka makhetlo a mararo ka boikemelo ka liphetho tse ts'oanang.
Bakeng sa lintlha tse ling mabapi le moralo oa thuto, bona Natural Portfolio Report Abstract e amanang le sengoloa sena.
Lintlha tse tala tsa MS tsa likarolo tse ntšitsoeng le data e tloaelehileng ea MS ea serum ea litšupiso li bonts'oa ho Supplementary Data 1 le Supplementary Data 2, ka ho latellana.Litlhaloso tsa tlhōrō bakeng sa likarolo tse fapaneng li hlahisoa ho Lintlha tsa Tlatsetso 3. Lethathamo la boitsebiso la LUAD TCGA le ka khoasolloa ho tsoa ho https://portal.gdc.cancer.gov/.Lintlha tse kentsoeng bakeng sa ho rala graph li fanoe mohloling oa data.Lintlha tsa mohloli li fanoe bakeng sa sengoloa sena.
National Lung Screening Study Group, joalo-joalo Ho fokotsa ho shoa ha kankere ea matšoafo ka tekanyo e tlaase ea computed tomography.Engelane Leboea.J. Med.365, 395–409 (2011).
Kramer, BS, Berg, KD, Aberle, DR le Prophet, PC Lung kankere ea ho hlahloba ho sebelisa tekanyo e tlaase ea helical CT: liphello tse tsoang ho National Lung Screening Study (NLST).J. Med.Screen 18, 109–111 (2011).
De Koning, HJ, et al.Ho fokotsa ho shoa ha mofetše oa matšoafo ka tlhahlobo ea volumetric CT tekong e sa reroang.Engelane Leboea.J. Med.382, 503–513 (2020).
Nako ea poso: Sep-18-2023